门静脉高压症脾大部切除后残脾基质金属蛋白酶及其抑制剂的变化

Changes of MMPs and tissue inhibitors of metalloproteinase in residual spleen after subtotal splenectomy for portal hypertension

摘要:

目的检测门静脉高压症脾大部切除后残脾组织和血清基质金属蛋白酶及其抑制剂的表达和水平，探讨脾纤维化的分子机制及保脾术的临床价值。方法选取门静脉高压脾肿大患者13 例。术中切取脾组织为巨脾组，术后8 年穿刺获取脾组织为残脾组，设外伤性脾破裂患者13 例为对照组。采用免疫组织化学法、实时荧光定量聚合酶链反应（qRT-PCR）及酶联免疫吸附测定（ELISA）检测脾脏组织和血清基质金属蛋白酶MMP-2、MMP-9、组织金属蛋白酶抑制物TIMP-1、TIMP-2 的表达和水平。结果3 组脾组织中，MMP-2、MMP-9、TIMP-1、TIMP-2 蛋白阳性表达主要分布在巨噬细胞内。残脾组和巨脾组脾组织MMP-2、MMP-9、TIMP-1 和TIMP-2 蛋白阳性表达率和基因表达与对照组比较，均差异有统计学意义（P <0.05），残脾组和巨脾组升高；残脾组和巨脾组之间比较则差异无统计学意义（P >0.05）。残脾组和巨脾组血清MMP-2、MMP-9、TIMP-1 和TIMP-2 水平与对照组比较，均差异有统计学意义（P <0.05），残脾组和巨脾组升高；残脾组和巨脾组之间比较差异无统计学意义（P >0.05）。结论门静脉高压症脾肿大患者脾大部切除术后残脾组织MMP-2、MMP-9、TIMP-1 及TIMP-2 蛋白和基因表达增加，血清蛋白水平升高，提示残脾组织基质金属蛋白酶和组织金属蛋白酶抑制物代谢失调。

Abstract:

Objective To measure the mRNA and protein levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) in residual splenic tissue and serum after subtotal splenectomy due to portal hypertension, and to investigate splenic fibrosis in molecular detail and the value of reserving splenic tissue during surgery. Methods Thirteen patients of splenomegaly due to portal hypertension who had subtotal splenectomy were enrolled in splenomegaly group. Those with spleen tissue puncture biopsy 8 years after splenectomy were selected into residual spleen group. Additionally, 13 traumatic spleen samples were collected for control group.Immunohistochemistry, PCR and ELISA were used to detect mRNA and protein levels of MMPs and TIMPs in the residual spleen samples and serum. Results In the residual spleen, splenomegaly and control groups, the positive expressions of MMP-2, MMP-9, TIMP-1 and TIMP-2 were densely distributed in macrophages. The positive expression rates of MMP -2, MMP -9, TIMP -1 and TIMP -2 proteins and mRNAs of the spleen tissues were significantly higher in the residual spleen and the splenomegaly groups compared with the control group (P < 0.05); however, there were no significant differences between the residual spleen group and the splenomegaly group (P >0.05). Significantly higher levels of serum MMP-2, MMP-9, TIMP-1 and TIMP-2 were observed in the residual spleen and the splenomegaly groups compared with the control group (P < 0.05); but there were no significant differences between the residual spleen and the splenomegaly groups (P > 0.05). Conclusions After subtotal splenectomy, MMP-2, MMP-9, TIMP-1 and TIMP-2 expressions are increased in the residual splenic tissue and serum of patients with portal hypertension. The results suggested that dysregulated MMPs and TIMPs expressions occur in the residual spleen after subtotal splenectomy due to portal hypertension.